Subtracted Restriction Fingerprinting (SRF) is a typing technique
recently described by Terletski
et al. (2003)
Check their article for more info.
SRF is a genotyping technique based on double endonuclease digestion of
genome. The endonucleases used in this simulation program are all
palinfromic, and they show no degenerated nucleotides. We will describe
briefly this technique:
SRF suggestion tool
- DNA is double digested with endonucleases.
- One endonuclease cleaves genome very frequently and the
other one fewer times.
- One restriction enzyme (RE1) allows performing a filling
reaction with digoxigenin labelled Uridine (Dig-dUTP) and the other
restriction enzyme allows a filling reaction with biotinilated Cytosine
- From the four types of fragments yielded after digestion,
RE1-RE2, RE2-RE1 and RE2-RE2 fragments (labelled in one or both ends
with biotin) are captured by streptavidin-coated magnetic beads
(they are substracted).
- Remaining RE1-RE1fragments (with are labelled in both end
with digoxigenin) will be
separated by electrophoresis, transfered by shouthern blot,
and bands pattern analysed.
The SRF suggestion tool
will provide researches with a list of endonuclease pairs suitable for
SRF fingerprinting of selected genome. Following the links, theoretical
electrophoretic pattern and identity of bands (sequence and ORF) may be